Glycolysis and oxidative phosphorylation are essential for purinergic receptor-mediated angiogenic responses in vasa vasorum endothelial cells.
Glycolysis and oxidative phosphorylation are essential for purinergic receptor-mediated angiogenic responses in vasa vasorum endothelial cells.
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DOI：
10.1152/ajpcell.00250.2016
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发表时间：
2017-01-01
期刊：
影响因子：
--
通讯作者：
Gerasimovskaya EV
中科院分区：
文献类型：
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作者：
Lapel M;Weston P;Strassheim D;Karoor V;Burns N;Lyubchenko T;Paucek P;Stenmark KR;Gerasimovskaya EV
血管生成是一个需要能量的过程;然而，细胞能量途径的作用及其通过细胞外刺激，特别是细胞外核苷酸的调节，仍然在很大程度上未被探索。使用糖酵解（2-脱氧葡萄糖）和氧化磷酸化（OXPHOS）（寡霉素，鱼藤酮，和FCCP）的代谢抑制剂，我们证明，糖酵解和OXPHOS都是血管内皮细胞（VVEC）的血管生成反应所必需的。用P2 R激动剂ATP和2-甲硫基腺苷二磷酸三钠盐（MeSADP）治疗，而不是P1受体激动剂腺苷，增加VVEC中的糖酵解活性（通过细胞外酸化速率和乳酸产生来测量）。糖酵解的刺激伴随着磷酸果糖激酶B3、己糖激酶（HK）和GLUT-1水平的增加，但不伴随乳酸脱氢酶。此外，细胞外ATP和MeSADP，并在较小程度上腺苷，增加VVEC的基础和最大耗氧率。与25 mM葡萄糖相比，当细胞在20 mM半乳糖和5 mM葡萄糖中培养时，这些作用增强。P2 R激动剂处理降低丙酮酸脱氢酶（PDH）-E1α的磷酸化，增加琥珀酸脱氢酶（SDH）、细胞色素氧化酶IV和F1 F0 ATP合酶β亚基的表达。此外，P2 R刺激瞬时升高线粒体Ca 2+浓度，这意味着线粒体参与VVEC血管生成激活。我们还证实了磷脂酰肌醇3-激酶和Akt通路在ATP刺激的VVEC中乳酸产生、PDH-E1α磷酸化以及HK、SDH和GLUT-1表达中的关键作用。总之，我们的研究结果表明，VVEC能量通路的嘌呤能和代谢调节是VV血管生成所必需的，并可能有助于肺动脉高压的病理性血管重塑。
Angiogenesis is an energy-demanding process; however, the role of cellular energy pathways and their regulation by extracellular stimuli, especially extracellular nucleotides, remain largely unexplored. Using metabolic inhibitors of glycolysis (2-deoxyglucose) and oxidative phosphorylation (OXPHOS) (oligomycin, rotenone, and FCCP), we demonstrate that glycolysis and OXPHOS are both essential for angiogenic responses of vasa vasorum endothelial cell (VVEC). Treatment with P2R agonists, ATP, and 2-methylthioadenosine diphosphate trisodium salt (MeSADP), but not P1 receptor agonist, adenosine, increased glycolytic activity in VVEC (measured by extracellular acidification rate and lactate production). Stimulation of glycolysis was accompanied by increased levels of phospho-phosphofructokinase B3, hexokinase (HK), and GLUT-1, but not lactate dehydrogenase. Moreover, extracellular ATP and MeSADP, and to a lesser extent adenosine, increased basal and maximal oxygen consumption rates in VVEC. These effects were potentiated when the cells were cultured in 20 mM galactose and 5 mM glucose compared with 25 mM glucose. Treatment with P2R agonists decreased phosphorylation of pyruvate dehydrogenase (PDH)-E1α and increased succinate dehydrogenase (SDH), cytochrome oxidase IV, and β-subunit of F1F0 ATP synthase expression. In addition, P2R stimulation transiently elevated mitochondrial Ca2+ concentration, implying involvement of mitochondria in VVEC angiogenic activation. We also demonstrated a critical role of phosphatidylinositol 3-kinase and Akt pathways in lactate production, PDH-E1α phosphorylation, and the expression of HK, SDH, and GLUT-1 in ATP-stimulated VVEC. Together, our findings suggest that purinergic and metabolic regulation of VVEC energy pathways is essential for VV angiogenesis and may contribute to pathologic vascular remodeling in pulmonary hypertension.